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Method for preparing cyclosporin A holoantigen

A complete antigen, cyclosporine technology, applied in the field of biotechnology and pharmacy, can solve the problems of high price, inconvenient patient medication, chronic rejection organ transplantation failure, etc., to save the synthesis process, shorten the photochemical reaction time, and shorten the reaction time. Effect

Inactive Publication Date: 2013-12-04
XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, in China, the automatic blood drug concentration monitor TDxFLx produced by Abbott of the United States is mostly used for blood drug concentration determination. The disadvantage is that it must purchase a special kit from Abbott of the United States. Because it is an exclusive production, the price is expensive, and it is expensive for patients. Great economic burden after surgery
Some patients reduce or give up the monitoring of CsA blood drug concentration midway due to economic reasons, resulting in CsA poisoning or chronic rejection organ transplantation failure in patients
In addition, at present, the use of TDxFLx is mostly concentrated in tertiary first-class hospitals. The blood drug concentration monitoring of CsA patients needs to go to these hospitals for drug concentration monitoring, which causes great inconvenience to patients.

Method used

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  • Method for preparing cyclosporin A holoantigen
  • Method for preparing cyclosporin A holoantigen
  • Method for preparing cyclosporin A holoantigen

Examples

Experimental program
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Embodiment 1

[0045] The preparation of embodiment 1 cyclosporin A complete antigen

[0046] Step 1: Weigh 240mg CsA and 50mg 4-benzoylbenzoic acid (BBa) with an electronic balance, and put them into a quartz cuvette (capacity 14mL), add 10mL tert-butanol with a pipette, and sonicate for 3min to fully dissolve CsA and 4-benzoylbenzoic acid (BBa);

[0047] Step 2: Prepare CsA incomplete antigen (CsA-BBa) by photochemical reaction method: After bubbling nitrogen gas into the cuvette in the above step 1 for 30±10min, turn on the ultraviolet light source and start to illuminate (the distance between the ultraviolet light source and the colorimetric dish 10±2cm), and at the same time unscrew the separating funnel above the cuvette (filled with a certain volume (300±50mL) of tert-butanol) to record the reaction time, so that tert-butanol drips slowly (1h before the drop rate 4 -8 drops / min, 1h 10-15 drops / min after the reaction), during the reaction process, continue to feed nitrogen gas, light ...

Embodiment 2

[0065] Identification of the cyclosporine A complete antigen prepared in Example 1:

[0066] 1. Identification of colorless solid powder product 2CsA incomplete antigen (CsA-BBa)

[0067] 1.1 TLC identification results

[0068] Chromatography (thin layer chromatography) of the product 2 in CHCl 3 / CH 3 OH=85 / 15 (volume ratio) binary expansion system, the test results confirmed: the Rf value of product 2 is 0.60, which is close to 0.58 measured in foreign patent literature, and it can be preliminarily judged that product 2 may be an incomplete antigen of CsA ( CsA-BBa).

[0069] 1.2 Melting point measurement results

[0070] The melting points of the reactant CsA, 4-benzoylbenzoic acid (BBa) and the product 2CsA incomplete antigen (CsA-BBa) were respectively measured with a melting point apparatus. The results are shown in Table 1:

[0071] Table 1. Melting points of CsA, BBa and product CsA-BBa

[0072]

[0073] It can be seen from Table 1 that the melting point of pr...

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Abstract

The invention provides a method for preparing cyclosporin A (CsA) holoantigen. According to the method, a high pressure mercury lamp is taken as a UV light source of a photochemical reaction, tert-butyl alcohol is taken as a reaction solvent, an intermediate CsA incomplete antigen is obtained by performing an addition reaction on 4-benzoylbenzoic acid and CsA under conditions of the photochemical reaction; the CsA incomplete antigen is dissolved in dimethyl formamide, and reacted with poly-L-lysine in water under the effect of 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride for synthesis of cyclosporin A holoantigen. The method is simple and feasibile and easy to operate, helps to shorten the photochemical reaction time of producing CsA incomplete antigen and substantially save the synthetic process of cyclosporin A holoantigen. ELISA reaction experiments and animal (mice) immunization experiments verify that the CsA holoantigen synthesized by the method has good specificity and immunogenicity, and the method is applicable to synthesis of CsA holoantigen.

Description

technical field [0001] The invention belongs to the fields of biotechnology and pharmacy, and in particular relates to a method for preparing cyclosporin A antigen. Background technique [0002] Cyclosporine A (CsA), as the main immunosuppressive drug for organ transplantation, plays an irreplaceable role in some fields of organ transplantation, and can effectively improve the survival time of transplanted organs. However, its therapeutic window is narrow, its bioavailability and pharmacokinetics vary greatly among individuals, and there are also certain differences in the body's sensitivity and tolerance to CsA. Therefore, long-term monitoring of blood drug concentration in patients is necessary to reduce the occurrence of toxic and side effects and prolong the survival time of transplanted organs. [0003] At present, in China, the automatic blood drug concentration monitor TDxFLx produced by Abbott of the United States is mostly used for blood drug concentration determin...

Claims

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Application Information

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IPC IPC(8): C08G69/48
Inventor 于丽秀华小黎陈东生吕永宁蔡雪峰伍三兰宋丽丽廖婧
Owner XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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